Anticorps Monoclonal anti-6*His, His-Tag
6*His, His-Tag Monoclonal Antibody for FC (Intra), IF
Hôte / Isotype
Mouse / IgG1
Réactivité testée
Protéine recombinante
Applications
IF, FC (Intra)
Conjugaison
CoraLite® Plus 647 Fluorescent Dye
CloneNo.
1B7G5
N° de cat : CL647-66005
Synonymes
Galerie de données de validation
Applications testées
| Résultats positifs en IF | cellules HEK-293 transfectées, |
| Résultats positifs en cytométrie | cellules HEK-293 transfectées, |
Dilution recommandée
| Application | Dilution |
|---|---|
| Immunofluorescence (IF) | IF : 1:250-1:1000 |
| Flow Cytometry (FC) | FC : 0.40 ug per 10^6 cells in a 100 µl suspension |
| It is recommended that this reagent should be titrated in each testing system to obtain optimal results. | |
| Sample-dependent, check data in validation data gallery | |
Applications publiées
| IF | See 1 publications below |
Informations sur le produit
CL647-66005 cible 6*His, His-Tag dans les applications de IF, FC (Intra) et montre une réactivité avec des échantillons Protéine recombinante
| Réactivité | Protéine recombinante |
| Hôte / Isotype | Mouse / IgG1 |
| Clonalité | Monoclonal |
| Type | Anticorps |
| Immunogène | Peptide |
| Nom complet | 6*His, His-Tag |
| Masse moléculaire calculée | 1 kDa |
| Symbole du gène | |
| Identification du gène (NCBI) | |
| Conjugaison | CoraLite® Plus 647 Fluorescent Dye |
| Excitation/Emission maxima wavelengths | 654 nm / 674 nm |
| Forme | Liquide |
| Méthode de purification | Purification par protéine G |
| Tampon de stockage | PBS avec glycérol à 50 %, Proclin300 à 0,05 % et BSA à 0,5 %, pH 7,3. |
| Conditions de stockage | Stocker à -20 °C. Éviter toute exposition à la lumière. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20oC Les 20ul contiennent 0,1% de BSA. |
Informations générales
Protein tags are a protein or peptide sequences located either on the C- or N- terminal of the target protein. His-tag is often used for affinity purification and binding assays. Expressed. The His-tag antibody is a useful tool for monitoring of the His-tagged proteins and recognizes His-tags placed at N-terminal, C-terminal, and internal regions of fusion proteins expressed in bacteria, insect, and mammalian cells. A His-tag (polyhistidine tag) consists of at least six histidine residues that are located at the N- or C-terminus of recombinant proteins. It is commonly used for affinity purification and protein binding experiments.
The recombinant protein has only 6 histidine residues as a tag. Is this sufficient to be detected by your antibody? Is the His-tag antibody able to detect N-terminal and C-terminal His tags?
The His-tag antibody was raised using a 6-His tag peptide as the immunogen, which is ~1 kDa. Some scientists use more than 6 histidine residues as a tag for recombinant proteins to increase affinity to metal ions. A 6-histidine tag is sufficient for specific recognition by the His-tag antibody. It is able to recognize both N-terminal and C-terminal His tags of recombinant proteins produced in various expression systems, including bacteria, yeasts, insect, and mammalian cells.
After purification, I can detect my protein by western blotting using the His-tag antibody but can also see additional bands running lower. What might they represent?
They may represent cleavage products of the recombinant protein. Unspecific protein cleavage can occur during protein production (e.g., in bacteria) or protein purification post the cell lysis steps. Changing expression conditions (temperature, time, induction methods, medium) and using protease inhibitors during purification helps to minimize protein degradation. Additional steps, such as ion exchange, size exclusion, or chromatography, allow isolation of the full-length product.
Can a His-tag antibody be used in applications other than western blotting?
Yes, this antibody has been successfully used for immunofluorescence (IF), immunoprecipitation (IP), and indirect ELISA analysis.
Protocole
| Product Specific Protocols | |
|---|---|
| IF protocol for CL Plus 647 6*His, His-Tag antibody CL647-66005 | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
| Species | Application | Title |
|---|---|---|
PLoS Pathog Novel secreted STPKLRR from Vibrio splendidus AJ01 promotes pathogen internalization via mediating tropomodulin phosphorylation dependent cytoskeleton rearrangement |
